A chemical and genetic approach together define the biological consequences of 3-methyladenine lesions in the mammalian genome.
DNA-damaging agents produce a plethora of cellular responses that include p53 induction, cell cycle arrest, and apoptosis. It is generally assumed that it is the DNA damage produced by these agents that triggers such responses, but there is limited direct evidence to support this assumption. Here, we used DNA alkylation repair proficient and deficient isogenic mouse cell lines to demonstrate that the signal to trigger p53 induction, cell cycle arrest, and apoptosis in response to alkylating agents does emanate from DNA damage. Moreover, we established that 3-methyladenine, a relatively minor DNA lesion produced by most methylating agents (which form mainly 7-methylguanine), can specifically induce sister chromatid exchange, chromatid and chromosome gaps and breaks, S phase arrest, the accumulation of p53, and apoptosis. This study was made possible by the generation of 3-methyladenine DNA glycosylase null mutant cells by targeted homologous recombination and by the chemical synthesis of a methylating agent that almost exclusively produces 3-methyladenine DNA lesions. The combined use of these two experimental tools has defined the biological consequences of 3-methyladenine, a DNA lesion produced by endogenous cellular metabolites, environmental carcinogens, and chemotherapeutic alkylating agents.
Engelward
B P
BP
Department of Cancer Cell Biology, Harvard School of Public Health, Boston, Massachusetts 02115, USA.
Allan
J M
JM
Dreslin
A J
AJ
Kelly
J D
JD
Wu
M M
MM
Gold
B
B
Samson
L D
LD
D000225:Adenine / Q000031:analogs & derivatives* / Q000378:metabolism; D000477:Alkylating Agents; D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002869:Chromosome Aberrations; D018736:DNA Adducts / Q000378:metabolism; D004249:DNA Damage; D045647:DNA Glycosylases; D019175:DNA Methylation; D004260:DNA Repair; D051379:Mice; D009699:N-Glycosyl Hydrolases / Q000172:deficiency*; D009429:Netropsin / Q000031:analogs & derivatives / Q000494:pharmacology; D012854:Sister Chromatid Exchange; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis
D000477:Alkylating Agents; D018736:DNA Adducts; D016159:Tumor Suppressor Protein p53; C110963:methyl lexitropsin; C025946:3-methyladenine; D009429:Netropsin; C021565:3-methyladenine-DNA glycosylase; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; D000225:Adenine